An enhanced visual detection assay for Listeria monocytogenes in food based on isothermal amplified peroxidase-mimicking catalytic beacon

2021 
Abstract Listeria monocytogenes is a kind of foodborne pathogen, which can cause meningitis, septicemia, gastroenteritis, and abortion with a fatality rate of up to 30%. Rapid, sensitive, and convenient detection of L. monocytogenes in food is an important strategy for controlling contamination and infection. In this study, an enhanced visual detection assay for L. monocytogenes based on an isothermal amplified peroxidase-mimicking catalytic beacon was developed. When L. monocytogenes existed, G-quadruplex sequences were amplified together with targeted gene sequence by polymerase spiral reaction (PSR). G-quadruplex adequately formed DNAzyme with hemin to have an enhanced peroxidase-like bioactivity with help of Nb. BbvCI nicking endonuclease, which could display significant colorimetric signals for indicating positive results qualitatively. The sensitivity test results determined that our assay enabled the quantitative detection of L. monocytogenes ranging from 10 to 105 CFU mL−1, as well as had a detection limit of 3.1 CFU mL−1, which was lower than previous visual detection assays for L. monocytogenes. Besides, it was notable that this assay could detect L. monocytogenes with the concentration of 2.0 lg CFU/g in artificially contaminated pork samples within about 4 h, with the advantages of high sensitivity, simple operation, and good visual signal output. We believed the improved visual detection assay based on isothermal amplified peroxidase-mimicking catalytic beacon showed great potential for food safety analysis.
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